Journal: Cell Biology and Toxicology

Article Title: CBX6 induces CD8 + T cell exhaustion and tumor development in esophageal squamous cell carcinoma through SMARCD1-mediated CCL8 secretion and lactate efflux

doi: 10.1007/s10565-025-10102-x

Figure Lengend Snippet: CBX6 induces CD8 + T cell exhaustion by promoting CCL8 secretion. A - C . Populations of Tex (CD8 + PD1 + TIM3 + ), Tpex (CD8 + PD1 + TIM3 – ), TNFα + CD8 + T, and IL-2 + CD8 + T cells in CD8 + T cells after a direct-contact co-culture with differentially treated mEC25 cells determined using FACS. D. Graphic presentation of the indirect, non-contact co-culture system of CD8 + T cells and differentially treated mEC25 cells. E. Populations of Tex (CD8 + PD1 + TIM3 + ) and Tpex (CD8 + PD1 + TIM3 – ) after the indirect co-culture determined using FACS. F. CM from differentially treated mEC25 cells was harvested to treat activated CD8 + T cells. G. Populations of Tex (CD8 + PD1 + TIM3 + ) and Tpex (CD8 + PD1 + TIM3 – ) cells after culturing in CM determined using FACS. H. Changes in cytokine-related gene expression in mEC25 cells after Cbx6 knockdown determined using RNA-seq analysis. I. Mouse recombinant proteins mIL6, mCCL8, mIL10, or control IgG were added to the non-contact co-culture system of CD8 + T cells and Cbx6-kd mEC25 cells. J. FACS analysis of the populations of Tex (CD8 + PD1 + TIM3 + ) and Tpex (CD8 + PD1 + TIM3 – ) cells after the supplementation of mIL6, mCCL8, mIL10, or control IgG. K. The non-contact co-culture system of CD8 + T cells and Cbx6-kd mEC25 cells were added with mCCL8 alongside DMSO or BMS-813160, an antagonist for the CCR8 receptor CCR5. L. FACS analysis of the populations of Tex (CD8 + PD1 + TIM3 + ) and Tpex (CD8 + PD1 + TIM3 – ) cells. M–O. mCCL8 alongside DMSO or BMS-813160 were added to the direct co-culture system of CD8 + T cells and Cbx6-kd mEC25 cells at different ratios, and then the populations of TNFα + CD8 + T, and IL-2 + CD8. + T cells were analyzed by FACS. P. Proliferation of mEC25 cells at different conditions. Six independent experiments were performed. Data from independent experiments are presented as dots in graphs. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Recombinant mouse Ccl8 (10 ng/mL, 250–15, PeproTech, USA) or the CCR5 antagonist BMS-813160 (HY-101482, 10 μM, MedChemExpress, USA) was supplemented to the experimental group.

Techniques: Co-Culture Assay, Gene Expression, Knockdown, RNA Sequencing, Recombinant, Control

Journal: Cell Biology and Toxicology

Article Title: CBX6 induces CD8 + T cell exhaustion and tumor development in esophageal squamous cell carcinoma through SMARCD1-mediated CCL8 secretion and lactate efflux

doi: 10.1007/s10565-025-10102-x

Figure Lengend Snippet: Cbx6 knockdown suppresses ESCC cell growth and promotes T cell function in vivo . A . mEC25 cells with or without Cbx6 knockdown were injected into immunocompromised BALB/c nude mice or immunocompetent C57BL/6 mice. B . Growth curve of mEC25 cells in nude mice. C. Representative images and weight of tumors in nude mice on day 21. D. Growth curve of mEC25 cells in C57BL/6 mice. E. Representative images and weight of tumors in C57BL/6 mice on day 21. F. Tumor remission rate [(Con-Cbx kd)/Con] in nude and C57BL/6 mice. G. FACS analysis of the proportions of immune-suppressive macrophages (CD11b + CD206 + ), Tregs (CD4 + Foxp3 + ), or MDSCs (CD11b + Ly6G + ) in tumor tissues of C57BL/6 mice. H. FACS analysis of the proportions of Tex and Tpex in tumor tissues of C57BL/6 mice. I-J. FACS analysis of the proportions of TNFα + and IL-2 + CD8 + T cells in tumor tissues of C57BL/6 mice. K. FACS analysis of the proportions of CCR5 + CD8 + T cells in tumor tissues of C57BL/6 mice. L. IHC analysis of the positive staining of C-Cas-3 in the tumor tissues of C57BL/6 mice. M. In another scenario, C57BL/6 mice received intra-tumoral administration of IgG or mCCL8 starting from day 7 after implantation of Cbx6-kd cells. N. Growth curve of mEC25 cells in mice. O. Representative images and weight of tumors in mice on day 21. P. FACS analysis of the populations of Tex and Tpex in tumor tissues. Q-R. FACS analysis of the proportions of TNFα + and IL-2 + CD8. + T cells in tumor tissues of mice. S. IHC analysis of the positive staining of C-Cas-3 in the tumor tissues. Each group contained six mice. Data from each experiment are presented as independent dots in graphs. * p < 0.05, *** p < 0.001, **** p < 0.0001

Article Snippet: Recombinant mouse Ccl8 (10 ng/mL, 250–15, PeproTech, USA) or the CCR5 antagonist BMS-813160 (HY-101482, 10 μM, MedChemExpress, USA) was supplemented to the experimental group.

Techniques: Knockdown, Cell Function Assay, In Vivo, Injection, Staining

Journal: Cell Biology and Toxicology

Article Title: CBX6 induces CD8 + T cell exhaustion and tumor development in esophageal squamous cell carcinoma through SMARCD1-mediated CCL8 secretion and lactate efflux

doi: 10.1007/s10565-025-10102-x

Figure Lengend Snippet: Smarcd1 overexpression induces CD8 + T cell exhaustion and enhances malignancy of mEC25 cells through CCL8 and MCT4. A - B . Enrichment of SMARCD1 in Slc16a3 or Ccl8 promoter in mEC25 cells with or without Cbx6 knockdown determined using ChIP-qPCR assays. C . mEC25 cells transfected with Cbx6 shRNA alongside Smarcd1 overexpression plasmid were co-cultured with activated CD8 + T cells, followed by addition of BMS-813160 or AZD0095 to the culture system. D . Relative survival of mEC25 cells after co-culture. E . FACS analysis of the populations of Tex and Tpex after co-culture. F . mEC25 cells transfected with Cbx6 shRNA alongside Smarcd1 overexpression plasmid were injected into C57BL/6 mice, followed by administration of BMS-813160 or AZD0095. G . Growth curve of mEC25 cells in mice. H - I . Representative images and weight of tumors in mice on day 21. P. FACS analysis of the populations of Tex and Tpex in tumor tissues. J . FACS analysis of the populations of Tex (CD8 + PD1 + TIM3 + ) and Tpex (CD8 + PD1 + TIM3 – ) in tumor tissues. K - L . FACS analysis of the proportions of TNFα + and IL-2 + CD8. + T cells in tumor tissues. M. IHC analysis of the positive staining of C-Cas-3 in the tumor tissues. For cellular experiments, six independent experiments were performed. For animal experiments, each group contained six mice. Data from each experiment are presented as independent dots in graphs. ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Recombinant mouse Ccl8 (10 ng/mL, 250–15, PeproTech, USA) or the CCR5 antagonist BMS-813160 (HY-101482, 10 μM, MedChemExpress, USA) was supplemented to the experimental group.

Techniques: Over Expression, Knockdown, ChIP-qPCR, Transfection, shRNA, Plasmid Preparation, Cell Culture, Co-Culture Assay, Injection, Staining

Journal: Cell Biology and Toxicology

Article Title: CBX6 induces CD8 + T cell exhaustion and tumor development in esophageal squamous cell carcinoma through SMARCD1-mediated CCL8 secretion and lactate efflux

doi: 10.1007/s10565-025-10102-x

Figure Lengend Snippet: Targeting Cbx6 enhances the therapeutic effect of anti-PD-1 therapy in mice bearing ESCC . A . Injection of luciferase-expressing mEc25 cells via tail vein into C57BL/6 mice, followed by treatment with Cbx6-siRNA and anti-PD-1 (IgG) alone or jointly on d 5, 7, 9, 11, and 13 post-injection. B - C . In vivo imaging analysis of fluorescence intensity to assess the growth and metastasis of mEc25 cells. D . Number of metastatic nodules in the lung tissues. E . FACS analysis of the proportion of Tex (CD8 + PD1 + TIM3 + ) and Tpex (CD8 + PD1 + TIM3 – ) in tumor tissues. F - G . FACS analysis of the proportion of TNFα + and IL-2 + CD8 + T cells in tumor tissues. H . FACS analysis of the proportion of CCR5 + CD8. + T cells in tumor tissues. I . Kaplan–Meier analysis of survival rate in each group of mice over 40 d. Each group contained six mice. Data from each experiment are presented as independent dots in graphs. * p < 0.05, *** p < 0.001, **** p < 0.0001

Article Snippet: Recombinant mouse Ccl8 (10 ng/mL, 250–15, PeproTech, USA) or the CCR5 antagonist BMS-813160 (HY-101482, 10 μM, MedChemExpress, USA) was supplemented to the experimental group.

Techniques: Injection, Luciferase, Expressing, In Vivo Imaging, Fluorescence

Journal: Cell Biology and Toxicology

Article Title: CBX6 induces CD8 + T cell exhaustion and tumor development in esophageal squamous cell carcinoma through SMARCD1-mediated CCL8 secretion and lactate efflux

doi: 10.1007/s10565-025-10102-x

Figure Lengend Snippet: CBX6 and SMARCD1 are potential clinical targets for ESCC treatment. A - B . IHC analysis of staining intensities of CBX6 and SMARCD1 in tumor and adjacent tissues of a TMA from 103 ESCC patients. C . Spearman correlation analysis of the staining intensities of CBX6 and SMARCD1. D - E . Patients with were divided into high-expression and low-expression groups based on the median staining intensity of CBX6 or SMARCD1, and PD-L1 staining intensity was detected in tumor tissues. F - G . The staining intensities of CBX6 and SMARCD1 in responders ( n = 11) and non-responders ( n = 6) following anti-PD-1 treatment. H - J . IHC analysis of CCL8, CCR5, and MCT4 staining intensities in tissues with high or low expression of CBX6 and SMARCD1. K - L . Kaplan–Meier analysis of the correlation between CBX6 and SMARCD1 staining intensity and OS or PFS of 89 patients. Data are presented as independent dots in graphs. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Recombinant mouse Ccl8 (10 ng/mL, 250–15, PeproTech, USA) or the CCR5 antagonist BMS-813160 (HY-101482, 10 μM, MedChemExpress, USA) was supplemented to the experimental group.

Techniques: Immunohistochemistry, Staining, Expressing

Journal: Water research

Article Title: Phytoextraction, phytotransformation and rhizodegradation of ibuprofen associated with Typha angustifolia in a horizontal subsurface flow constructed wetland.

doi: 10.1016/j.watres.2016.06.049

Figure Lengend Snippet: Fig. 4. Occurrence of ibuprofen carboxylic acid (CBX IBP) in the interstitial water samples collected along the flow path in the planted and unplanted beds. The values below quantification limit (1.38 mg L1) are plotted with the value of 1.37 mg L1 1/3L and 2/3L are the locations at the 1/3 length and 2/3 length of the treatment section along the flow path, respectively. Sampling campaigns: n ¼ 36.

Article Snippet: High purity ( 98%) ibuprofen carboxylic acid (CBX IBP) was provided by Toronto Research Chemicals (Canada).

Techniques: Sampling

Journal: Water research

Article Title: Phytoextraction, phytotransformation and rhizodegradation of ibuprofen associated with Typha angustifolia in a horizontal subsurface flow constructed wetland.

doi: 10.1016/j.watres.2016.06.049

Figure Lengend Snippet: Fig. 5. Mean concentrations of ibuprofen (IBP), ibuprofen carboxylic acid (CBX IBP), 2- hydroxy ibuprofen (2-OH IBP) and 1-hydroxy ibuprofen (1-OH IBP) in the lamina and sheath tissues of Typha angustifolia collected at 1/3L (a) and 2/3L (b) in the planted bed. 1/3L and 2/3L are the locations at the 1/3 length and 2/3 length of the treatment section along the flow path, respectively. Sampling campaigns: n ¼ 36. The normalized IBP concentrations calculated from the concentrations of IBP metabolites are illus- trated for the two sampling locations. *: p < 0.05 (t-test).

Article Snippet: High purity ( 98%) ibuprofen carboxylic acid (CBX IBP) was provided by Toronto Research Chemicals (Canada).

Techniques: Sampling